Synthetic biology has emerged as a useful approach to decoding fundamental laws underlying biological control. Recent efforts have produced many systems and approaches and generated substantial insights on how to engineer biological functions and efficiently optimize synthetic pathways.
Despite efforts and progresses, current approaches to perform such engineering are often laborious, costly and difficult. Challenges still remain in developing engineering-driven approaches and systems to accelerate the design-build-test cycles required for reprogramming existing biological systems, constructing new biological systems and testing genetic circuits for transformative future applications in diverse areas including biology, engineering, green chemistry, agriculture and medicine.
An in vitro transcription-translation (TX-TL) system (Shin & Noireaux, 2012; Sun et al., 2013) has been developed which allows for the rapid prototyping of genetic constructs (Sun, et al., 2014) in an environment that behaves similarly to a cell (Niederholtmeyer et al., 2015; Takahashi et al., 2015). One of the main purposes of working in vitro is to be able to learn or characterize a circuit for future implementation in vivo (Chappell et al., 2013; Niederholtmeyer et al., 2015). However, there are no easy ways to convert deoxyribonucleic acid (DNA), which was created primarily for in vitro testing, to make the DNA compatible for the in vivo environment when implemented on plasmid. In specific, origins of replication need to be in compatible families, and antibiotic resistance markers need to be varied per plasmid. Thus, a need exists for new techniques that can overcome these challenges.